The purpose of this lab is to extract and quantify some major cellular components, which are pigments contained in chloroplasts, DNA, RNA, and proteins. Euglena gracilis is a single-celled eukaryote that has the contains all of these components and so they were the model organism in this experiment. The purpose of this experiment was achieved by utilizing different reactions that are able to extract specific cellular components. The concentrations of these components were measured using absorbance and these values were used to determine the amount of components present in an individual Euglena cell. Additionally, the morphology of the cells was noted. The ratios of the cellular components of the Euglena suspension was compared to the standardized …show more content…
With each reaction, the solution was vortex and centrifuged. The supernatant was collected in a separate tube and the pellet was reacted with 5 ml of acetone. In this experiment, the first supernatant of the three was accidently discarded, therefore only two of the three supernatants were collected for total volume of 10 ml of supernatant A, rather than 15 ml. Using the UltraSpec 3000, an absorbance spectrum was measured at various wavelengths from 400-700 nm (Figure 1). 80% acetone was used as a blank. Since the some of the sample used for these readings was lost, no additional dilutions were needed. Chlorophyll Determination: The absorbance at 652 nm was measured to determine the total number of chlorophyll a and b in the supernatant. At a wavelength of 652 nm, the two have the same absorbance and carotenoids do not absorb the light. The absorbance was 0.110 A, which was consistent with the standard. The concentration of the total chlorophyll was determined using Beer’s Law: mg/ml. Since the concentration of the suspension was approximately 1.8X106 mg/ml, there are approximately: mg of chlorophyll a and b in each cell of the Euglena suspension. Carotenoid